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Optimized amplification of BK polyomavirus in urine
Elizabeth A Odegard1, Heidi L Meeds1, Steven B Kleiboeker2
1Division of Digestive Diseases, University of Cincinnati College of Medicine, Cincinnati, OH, United States.
Journal of Virological Methods
|October 10, 2021
Summary
BK polyomavirus (BKPyV) DNA can be extracted from urine for next-generation sequencing. This allows for accurate BKPyV genotyping and analysis of mutations linked to disease in immunocompromised patients.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- BK polyomavirus (BKPyV) is a common virus causing asymptomatic infections in most people.
- In immunocompromised individuals, BKPyV can shed at high levels (up to 10^9 copies/mL) in urine.
- High viral loads in urine present an opportunity for detailed genetic analysis.
Purpose of the Study:
- To describe methods for extracting BKPyV DNA from urine.
- To outline amplification techniques for preparing urine samples for next-generation sequencing (NGS).
- To enable accurate BKPyV genotyping and identification of pathogenic mutations.
Main Methods:
- DNA extraction from urine samples.
- Polymerase chain reaction (PCR) amplification of viral DNA.
- Preparation of libraries for next-generation sequencing.
Main Results:
- Successful extraction of BKPyV DNA from urine samples.
- Optimized amplification protocols for sequencing library preparation.
- Demonstrated feasibility of using urine for BKPyV genetic characterization.
Conclusions:
- Urine is a suitable sample source for BKPyV DNA extraction and NGS.
- These methods facilitate accurate BKPyV genotyping and mutation analysis.
- Understanding BKPyV genetic diversity can inform clinical management.
